BACKGROUNDOsteosarcoma is characterized by high neovascularization and a high propensity for metastasis through bloodstream. This study was to examine whether there is evidence for vasculogenic mimicry in osteosarcoma and to illustrate mechanism of tumor blood vessels formation in osteosarcoma.

METHODSOsteosarcoma cell lines (U-2OS) were tested for their ability to form tubular networks in three-dimensional culture containing type I collagen. The structures of the tubular networks were observed with phase contrast microscope and transmission electron microscope (TEM). Morphometric studies using hematoxylin and eosin (HE) stain and CD31 immunohistochemical stain to show tumor-lined channels in human osteosarcoma were also performed.

RESULTSObservation with light microscope and TEM showed that highly aggressive osteosarcoma cell lines (U-2OS) formed networks containing channels when grown in three-dimensional culture containing type I collagen, in the absence of endothelial cells or fibroblasts. Morphometric observation using HE stain and CD31 immunohistochemical stain showed that tumor cell-lined channels were also detected in vivo in osteosarcoma; by comparison, all vascular areas in the pedicle of osteochondroma or outside osteochondroma were endothelial-lined.

CONCLUSIONThese observations strongly suggest that aggressive osteosarcoma cells may generate vascular channels that facilitate tumor perfusion independent of tumor angiogenesis and have the ability of vasculogenic mimicry.

Bone Neoplasms ; pathology ; ultrastructure ; Cell Line, Tumor ; Collagen ; biosynthesis ; Humans ; Immunohistochemistry ; Neovascularization, Pathologic ; pathology ; Osteosarcoma ; pathology

Objective To evaluate the association between total fluid intake and the time of urination per day and the risk of bladder cancer. Methods A population-based case-control study was conducted in urban Shanghai, China, during January 1996 to December 1998. The study included 608 incident cases of bladder cancer and 607 age- and sex-matched controls. Unconditional logistic regression models were used to estimate the odds ratios(ORs)and their corresponding 95%confidence intervals(95%CIs)for bladder cancer associated with frequency of urination, after adjusted for age, gender, smoking status, history of occupation with high risk, history of bladder infections, body mass index and other confounding factors. The level of statistical significance was set at 0.05(two-sided). Results No significant trend was observed for the association between total fluid intake, time of nighttime urination and the risk of bladder cancer. Increasing time of urination during daytime was associated with decreased risk of bladder cancer(P for trend=0.014). ORs(95%CIs)for subjects who voided 4 times, 5 times and 6 or more times per day[0.72(0.49-1.05),0.60(0.41-0.87)and 0.62(0.43-0.90), respectively], when compared with those with less than 4times per day after adjustment of confounding factors. Data showed that smokers and nonsmokers who voided at least 6 times per day had the ORs of 0.72(95%CI: 0.45-1.15)and 0.46(95%CI:0.25-0.87)when compared to their counterparts who voided 3 times or less per day during the daytime. Subjects who urinated at least 6 times per day and consumed more than 1500 ml of total fluid per day experienced a significant 57% reduction in risk compared to subjects who urinated 3 times or less and consumed less than 750 ml of total daily fluid intake. Conclusion Increased urination frequency and total fluid intake, especially among those who never smoked might be associated with a reduced risk of bladder cancer.

OBJECTIVETo observe and assess the immunosuppressive effect of applying bailing capsule (BLC, a dry powder preparation of Cordyceps sinensis mycelia), after renal transplantation, its influence on other systems of organism, and to explore the possible therapeutic mechanism.

METHODSOne hundred and twenty-one recipients of renal homo-allograft were randomly divided into two groups. The 64 cases in Group A was treated with cyclosporin A (Cs A) + prednisone (pred) + azathioprine (Aza), the 57 in Group B treated with Cs A + pred + BLC. They were followed-up for 1-2 year by checking up blood routine, urine routine, liver and renal function, blood electrolytes, glucose and lipids, and uric acid for 2 times every week in the first month after transplantation, followed by proper re-examination of these items according to various condition.

RESULTSThere was no significant difference between the two groups in aspects of graft survival rate, occurrence of reject reaction, renal function recovery, blood electrolytes and blood glucose levels. However, as compared with Group A, in Group B, levels of urinary erythrocytes and leucocytes, blood alanine transaminase (ALT), aspartate aminotransferase (AST), total cholesterol, uric acid as well as the incidence of infection were significantly lower, and blood high density lipoprotein, serum total protein, albumin, RBC and WBC count were significantly higher.

CONCLUSIONBLC could effectively prevent the reject response after renal transplantation, protect renal and liver function, stimulate hemopoietic function, improve hypoproteinemia and hyperlipidemia, reduce the infection, etc., therefore, it is an ideal immunosuppressor after organ transplantation.

Adult ; Capsules ; Cordyceps ; Cyclosporine ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Glomerulonephritis ; drug therapy ; surgery ; Graft Rejection ; prevention & control ; Humans ; Immunosuppressive Agents ; therapeutic use ; Kidney Transplantation ; Male ; Phytotherapy ; Postoperative Period ; Prednisone ; therapeutic use

OBJECTIVETo rapidly and sensitively detect the four virulence-associated factors of Streptococcus suis, a multiplex PCR was developed.

METHODSIn the process of this reaction, four distinct DNA targets were amplified. One target was based on the serotype 2 (and 1/2) specific cps gene and the others were based on Streptococcus suis mrp, epf (epf*) and sly gene, encoding the MRP, EF(EF*) and Sly proteins of Streptococcus suis. 72 isolates, which including 48 strains of Streptococcus suis and 24 strains of negative control, and 49 clinical specimens were detected by the multiplex PCR assay.

RESULTSAll PCR products were detected by electrophoresis on 1.2% agarose gels. With the 48 Streptococcus suis strains, the positive detection rates of cps2+, mrp+, epf+, epf*+ and sly+ were 16/48, 14/48, 12/48, 3/48 and 26/48,respectively. The results were confirmed by bacteriological examination. There were no specific amplification products including 49 clinical specimens and 24 negative control strains.

CONCLUSIONThe results demonstrated that multiplex PCR was a highly specific and sensitive diagnostic tool for the detection of virulence-associated factors of streptococcus suis.

Bacterial Proteins ; genetics ; Polymerase Chain Reaction ; methods ; Streptococcus suis ; genetics ; pathogenicity ; Virulence Factors ; genetics

OBJECTIVETo investigate the chronic effects of intracoronary autologous bone marrow mononuclear cell (BM-MNCs) transplantation in patients with refractory heart failure (RIHF) after myocardial infarction.

METHODSThirty patients with RIHF (LVEF < 40%) were enrolled in this nonrandomized study, autologous BM-MNCs (5.0 +/- 0.7) x 10(7) were transplanted with via infarct-related coronary artery in 16 patients and 14 patients received standard medical therapy served as control. Baseline and follow up evaluations included complete clinical evaluations, plasma BNP, ANP, ET-1 measurements, echocardiography, PET, and Holter monitoring.

RESULTSBaseline characteristics were similar between the 2 groups. There were no major periprocedural complications. One patient developed ventricular premature contractions during cell infusion for several seconds and recovered spontaneously. Compared to pre-transplantation, plasma BNP and ET-1 significantly decreased and plasma ANP significantly increased at 7 days post transplantation; 6 minutes walking distance increased from (72.1 +/- 31.5) to (201.6 +/- 23.3) m (P < 0.01), LVEF increased 9.9% (P < 0.001) and FDG-PET revealed vital myocardium area increased (10.3 +/- 3.4)% (P < 0.01) at 3 months after BM-MNCs transplantation. At 6 months follow up, the NYHA class improved from (3.4 +/- 0.1 to 2.4 +/- 0.2, P < 0.001) and no patient died and 1 patient rehospitalized due to lower extremities edema. In control group, LVEF decreased 7.2% compared to baseline (P < 0.001) and was significantly lower than transplantation group at 3 months (P < 0.001). At 6 months follow up, the NYHA class increased from (3.5 +/- 0.1 to 3.9 +/- 0.1, P < 0.05), 2 patients died and 10 patients rehospitalized due to aggravated heart failure.

CONCLUSIONPresent study demonstrates that intracoronary transplantation of autologous BM-MNCs is safe and effective for treating patients with RIHF after myocardial infarction.

Bone Marrow Transplantation ; Coronary Vessels ; surgery ; Follow-Up Studies ; Heart Failure ; complications ; Humans ; Mesenchymal Stem Cell Transplantation ; Monocytes ; transplantation ; Myocardial Infarction ; surgery ; Myocardial Ischemia ; complications ; Transplantation, Autologous

Aged ; Blood Glucose ; metabolism ; Female ; Humans ; Hyperglycemia ; epidemiology ; etiology ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Postoperative Period ; Retrospective Studies

AIM:To observe the changes of autophagy-related indexes during endoplasmic reticulum stress (ERS) induced by dithiothreitol (DTT) and its effect on apoptosis in human normal hepatocytes.METHODS:LO2 cells were treated with DTT at 2.0 mmol/L for 0, 6, 12 and 24 h to induce ERS.The expression of glucose-regulated protein 78 (GRP78) , protein kinase R-like endoplasmic reticulum kinase (PERK) , activating transcription factor 4 (ATF4) , C/EBP homologous protein (CHOP) , autophagy-related gene 12 (Atg12) , autophagy-related gene 5 (Atg5) and microtubule-associated protein 1 light chain 3 (LC3) at mRNA and protein levels was determined by real-time PCR and Western blot.The apoptosis was analyzed by flow cytometry.The formation of autophagosomes was observed under transmission electron microscope.After the LO2 cells were pretreated with rapamycin at 400 nmol/L for 1 h and treated with DTT at 2.0mmol/L for 24 h, the effect of rapamycin pretreatment on the apoptosis was analyzed by flow cytometry.RESULTS:After treatment with DTT at 2.0 mmol/L for 6, 12 and 24 h, the mRNA and protein levels of GRP78, PERK, ATF4, CHOP, Atg12, Atg5 and LC3 in the LO2 cells were significantly higher than those in 0 h group (P<0.05).At the same time, the ratio of LC3Ⅱ/LC3Ⅰwas also increased after DTT treatment (P<0.05).Observation under transmission electron microscope showed that autophagosomes were found in the LO2 cells treated with DTT for 6, 12 and 24 h.After DTT treatment for 6, 12 and 24 h, the apoptosis rate of LO2 cells was significantly higher than that in DTT 0 h group, while the apoptosis induced by DTT was significantly decreased after rapamycin pretreatment (P<0.05).CONCLUSION:ERS induces autophagy and rapamycin pretreatment alleviates the apoptosis of LO2 cells to some extent.

AIM:To investigate the effect of SET7/9 (SET domain containing 7/9) -mediated endoplasmic reticulum stress (ERS) on protein kinase R-like endoplasmic reticulum kinase (PERK) signaling pathway, and to explore the mechanisms of arsenic-induced hepatocyte apoptosis.METHODS:Human liver LO2 cells were divided into control group, arsenic poisoning model group, negative transfection group and SET7/9 siRNA transfection group.The apoptosis of the LO2 cells in each group was analyzed by flow cytometry.The protein levels of SET7/9, glucose-regulated protein 78 (GRP78) , PERK and p-PERK in the LO2 cells of each group were observed by Western blot.RESULTS:Inhibition of SET7/9 expression reduced the apoptotic rate of arsenic-induced LO2 cells.Arsenic exposure increased the expression of SET7/9 in the LO2 cells.Arsenic exposure increased the protein levels of GRP78 and p-PERK in the LO2 cells, but decreased the protein levels of GRP78 and p-PERK after transfection with SET7/9 siRNA (P<0.05).CONCLUSION:Arsenic exposure induces hepatocyte apoptosis by increasing SET7/9 to activate ERS by PERK signaling pathway.

Objective To investigate the effect of replacing a smaller-size catheter after tubularized incised plate (TIP) urethroplasty on postoperative urethral complications.Methods The data of 116 hypospadias patients underwent TIP urethroplasty performed by the same urologist in our hospital from January 2014 to December 2017 were retrospectively analyzed.The age of patients ranged from 0.5-15.4 years,with the median age of 1.5 years.Meatal location was distal in 47 (39.5％),midshaft in 62 (52.1％) and proximal in 10(8.4％) patients.Unhealthy urethral plate occurred in 49 (42.2％) patients.F8 catheter was used for urethroplasty in 92 (79.3 ％) patients,F10 in 17 (14.7％),and F12 in 7 (6.0％).According to the pattern of urinary diversion,patients were divided into two groups.Conventional catheter group in 56 patients (group A):the catheter was chosen in as large size as possible for urethroplasty without tension.Replace a smaller-size catheter group in 60 patients (group B):replace a smaller-size catheter after urethroplasty on the basis of group A.There were no statistically significant differences in age,meatal location and catheter size between the two groups (P ＞ 0.05).Results The mean follow up was 23 months (range 6-66 months).There were 73(62.9％) cases of orifice overflow during indwelling catheterization.There were 5 (4.3％) patients needed further reoperation result from postoperative urethral complications,including 4 cases of fistula and 1 case of meatal stenosis.The process of replacing a smaller-size catheter after urethroplasty in group B was simple and without adverse reactions.There were 15 (26.8％) cases of orifice overflow in group A and 58 (96.7％) in group B.There were statistically significant differences in orifice overflow between the two groups (P ＜ 0.05).There were 5 (8.9％) cases of postoperative urethral complications in group A and 0 case in group B.There were statistically significant differences in postoperative urethral complications (P ＜ 0.05).Conclusions The process of replacing a smaller-size catheter after TIP urethroplasty in was simple and subsequently contributed to less postoperative urethral complications.The results of replacing a smaller-size catheter after TIP urethroplasty were superior to that of no replacing conventionally.

OBJECTIVETo explore the pathogenesis of tumors by blocking the normal differentiation process of stem cells.

METHODSBone marrow mesenchymal stem cells (BMSCs) from rats were isolated, cultured and purified by whole bone marrow adherence method. The rat BMSCs were induced to differentiate into adipocytes with dexamethasone, insulin and indomethacin. Blockage of the differentiation process was induced by 3-methylcholanthrene (3-MC).

RESULTSThe differentiation experiment showed that at 30 days after the induction, oil red O staining-positive cells occurred with increased intracytolasmic lipid droplets, characteristic for adipocytes. The differentiation blockage experiment showed that at 30 days after induction, the deposits of oil red O staining-cytoplasmic lipid droplets was significantly reduced, indicating that the blocked cells were adipocytes, but not fully differentiated. Morphological identification showed that cell contact inhibition disappeared, abnormal cell nuclei, increased number of micronucleus aberration and karyotype abnormalities, indicating that malignant transformation of the stem cells occurred after the differentiation blockage.

CONCLUSIONSThe results of this study show a blockage of the differentiation of that stem cells at the intermediate phase, and a tendency of malignant transformation of the stem cells. The results of our study provide new evidence that cancer stem cells may be originated by suppression of stem cell differentiation.

Adipocytes ; cytology ; Animals ; Bone Marrow Cells ; cytology ; drug effects ; Cell Differentiation ; drug effects ; Cell Transformation, Neoplastic ; Cells, Cultured ; Dexamethasone ; pharmacology ; Drug Combinations ; Female ; Indomethacin ; pharmacology ; Insulin ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Methylcholanthrene ; pharmacology ; Rats ; Rats, Wistar